摘要
目的建立基于阳离子交换固相萃取(SPE)小柱净化,液相色谱-串联质谱(LC-MS/MS)检测腌制河豚鱼中河豚毒素(TTX)的方法。方法粉碎均匀的腌制河豚鱼样品,采用0.5%乙酸/50%甲醇/水超声提取,提取液经阳离子交换SPE小柱净化,0.3%盐酸/50%乙腈/水洗脱,洗脱液经氨水中和后采用XBridgeTM BEH Amide色谱柱(150 mm×3.0 mm,1.7μm)分离,采用串联质谱多反应监测模式测定。结果TTX的基质效应为85.7%~92.4%,采用优化后的SPE方法净化,基质抑制效应得到有效控制。样品中TTX在2.0~4000μg/kg范围内线性关系良好,相关系数(r2)为0.9992;方法检出限和定量限分别为1.0μg/kg和2.0μg/kg。在2.0、200、2200μg/kg水平加标,平均回收率为81.2%~96.5%,相对标准偏差为4.3%~7.5%。质控样品中TTX的日内准确度为84.4%~95.6%,精密度为4.9%~5.8%;日间准确度为86.1%~94.9%,精密度为5.5%~8.5%。采用本方法测定38份市售腌制河豚鱼制品,TTX检出率为60.5%。结论本研究建立的方法可以实现腌制河豚鱼中TTX的准确定量测定。
Objective To develop a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method with cation exchange-based solid phase extraction(SPE)for determination of tetrodotoxin(TTX)in salted pufferfish.Methods Evenly crushed salted pufferfish samples were subjected to ultrasound-assisted extraction with 0.5%acetic acid/50%methanol/water.The extract was cleaned with cation exchange-based SPE cartridge and eluted with 0.3%hydrochloric acid and 50%acetonitrile/water.The eluent was neutralized with ammonia and separated with a Waters XBridgeTM BEH Amide column(150 mm×3.0 mm,1.7μm),and determined using LC-MS/MS in a multiple reaction monitoring mode.Results The matrix effects of TTX were 85.7%-92.4%,and the matrix suppression effect was under effective control following clean-up procedures using the optimized SPE method.The TTX showed a good linear relationship at the range of 2.0 to 4000μg/kg,with a correlation coefficient(r2)of 0.9992.The limits of detection and quantitation for TTX in sample matrix were 1.0μg/kg and 2.0μg/kg,respectively.The mean spiked-recovery rates were 81.2%to 96.5%at spiked amounts of 2.0,200μg/kg and 2200μg/kg,with relative standard deviations(RSDs)of 4.3%to 7.5%.The intraday accuracy and precision of TTX were 84.4%to 95.6%and 4.9%to 5.8%in quality control samples,and the interday accuracy and precision of TTX were 86.1%to 94.9%and 5.5%to 8.5%in quality control samples.The detection of TTX was 60.5%in 38 market-sold salted pufferfish products using the established LC-MS/MS method.Conclusion The established LC-MS/MS method is effective for accurate quantitative determination of TTX in salted pufferfish.
作者
俞莎
徐美佳
徐小民
YU Sha;XU Meijia;XU Xiaomin(Department of Physicochemical and Toxicology,Zhejiang Provincial Center for Disease Control and Prevention,Hangzhou,Zhejiang 310051,China)
出处
《预防医学》
2023年第7期640-644,共5页
CHINA PREVENTIVE MEDICINE JOURNAL
基金
浙江省医药卫生科技计划(2021KY615)。
关键词
腌制河豚鱼
河豚毒素
固相萃取
阳离子交换
液相色谱-串联质谱
salted pufferfish
tetrodotoxin
solid phase extraction
cation exchange
liquid chromatography-tandem mass spectrometry
